LncRNA lnc-ISG20 promotes renal fibrosis in diabetic nephropathy by inducing AKT phosphorylation through miR-486-5p/NFAT5.

Long non-coding RNA (lncRNA) lnc-ISG20 has been found aberrantly up-regulated in the glomerular in the patients with diabetic nephropathy (DN). We aimed to elucidate the function and regulatory mechanism of lncRNA lnc-ISG20 on DN-induced renal fibrosis. Expression patterns of lnc-ISG20 in kidney tissues of DN patients were determined by RT-qPCR. Mouse models of DN were constructed, while MCs were cultured under normal glucose (NG)/high glucose (HG) conditions. The expression patterns of fibrosis marker proteins collagen IV, fibronectin and TGF-ß1 were measured with Western blot assay. In addition, the relationship among lnc-ISG20, miR-486-5p, NFAT5 and AKT were analysed using dual-luciferase reporter assay and RNA immunoprecipitation. The effect of lnc-ISG20 and miR-486/NFAT5/p-AKT axis on DN-associated renal fibrosis was also verified by means of rescue experiments. The expression levels of lnc-ISG20 were increased in DN patients, DN mouse kidney tissues and HG-treated MCs. Lnc-ISG20 silencing alleviated HG-induced fibrosis in MCs and delayed renal fibrosis in DN mice. Mechanistically, miR-486-5p was found to be a downstream miRNA of lnc-ISG20, while miR-486-5p inhibited the expression of NFAT5 by binding to its 3'UTR. NFAT5 overexpression aggravated HG-induced fibrosis by stimulating AKT phosphorylation. However, NFAT5 silencing reversed the promotion of in vitro and in vivo fibrosis caused by lnc-ISG20 overexpression. Our collective findings indicate that lnc-ISG20 promotes the renal fibrosis process in DN by activating AKT through the miR-486-5p/NFAT5 axis. High-expression levels of lnc-ISG20 may be a useful indicator for DN.

Exosomal microRNA-16-5p from human urine-derived stem cells ameliorates diabetic nephropathy through protection of podocyte.

Diabetic nephropathy (DN) remains one of the severe complications associated with diabetes mellitus. It is worthwhile to uncover the underlying mechanisms of clinical benefits of human urine-derived stem cells (hUSCs) in the treatment of DN. At present, the clinical benefits associated with hUSCs in the treatment of DN remains unclear. Hence, our study aims to investigate protective effect of hUSC exosome along with microRNA-16-5p (miR-16-5p) on podocytes in DN via vascular endothelial growth factor A (VEGFA). Initially, miR-16-5p was predicated to target VEGFA based on data retrieved from several bioinformatics databases. Notably, dual-luciferase report gene assay provided further verification confirming the prediction. Moreover, our results demonstrated that high glucose (HG) stimulation could inhibit miR-16-5p and promote VEGFA in human podocytes (HPDCs). miR-16-5p in hUSCs was transferred through the exosome pathway to HG-treated HPDCs. The viability and apoptosis rate of podocytes after HG treatment together with expression of the related factors were subsequently determined. The results indicated that miR-16-5p secreted by hUSCs could improve podocyte injury induced by HG. In addition, VEGA silencing could also ameliorate HG-induced podocyte injury. Finally, hUSC exosomes containing overexpressed miR-16-5p were injected into diabetic rats via tail vein, followed by qualification of miR-16-5p and observation on the changes of podocytes, which revealed that overexpressed miR-16-5p in hUSCs conferred protective effects on HPDCs in diabetic rats. Taken together, the present study revealed that overexpressed miR-16-5p in hUSC exosomes could protect HPDCs induced by HG and suppress VEGFA expression and podocytic apoptosis, providing fresh insights for novel treatment of DN.

[Retracted] MicroRNA­21 promotes the progression of peritoneal fibrosis through the activation of the TGF­ß/Smad signaling pathway: An in vitro and in vivo study.

Int J Mol Med 41: [Related article:] 1030­1038, 2018; DOI: 10.3892/ijmm.2017.3268. An interested reader drew to our attention the fact that the western blots featured in Fig. 2B in the above article contained duplicated data: The data shown for the TGF­ß and vimentin protein bands were apparently identical; furthermore, there was a strong likelihood that the protein bands featured for the ZO­1 and SMAD­3 experiments were also the same, but flipped horizontally relative to the other. Following an investigation, the Journal was able to confirm that this duplication of the research data had probably occurred. On those grounds, the Editor of International Journal of Molecular Medicine has decided that the above paper should be retracted. We were unable to make contact with the authors of the article published in International Journal of Molecular Medicine, despite every effort to do so. The Editor deeply regrets any inconvenience that this retraction has caused to the the readership of the Journal.

Rhein inhibits malignant phenotypes of human renal cell carcinoma by impacting on MAPK/NF-κB signaling pathways.

BACKGROUND: Rhein, an anthraquinone derivative of rhubarb, is traditionally used in Chinese herbal medicine. Now emerging studies suggest its antitumor properties in many human cancers. The present study aims to investigate the antitumor role of Rhein and its possible mechanism in human renal cell carcinoma (RCC). MATERIALS AND METHODS: Three RCC cell lines (A489, 786-O and ACHN) were used as the cell models. We applied CCK-8, cell counting, colony formation, wound healing and Transwell assays to assess the antitumor roles of Rhein in RCC cells in vitro. The therapeutic efficacy of Rhein was further evaluated by intraperitoneal administrations in tumor formation of mice. Western blot was used to investigate the underlying mechanisms of action of Rhein. RESULTS: Rhein inhibited RCC cell proliferation in a dose- and time-dependent manner. It also suppressed RCC cell migration and invasion in vitro. Moreover, Rhein was able to inhibit tumor growth in nude mice by intraperitoneal administration in vivo. Mechanistically, the protein levels of phosphorylated MAPK (mitogen-activated protein kinase, extracellular signal-regulated kinase and c-Jun N-terminal kinase), phosphorylated Akt and two targets of NF-κB (nuclear factor kappa-light-chain enhancer of activated B cells) pathway, matrix metalloproteinase 9 and CCND1 were all markedly reduced by Rhein treatment. CONCLUSION: Rhein processed the antitumor effects in RCC cells by inhibiting cell proliferation, migration and invasion, and these tumor-suppressing functions might be mediated by MAPK/NF-κB signaling pathways.

MicroRNA-21 promotes the progression of peritoneal fibrosis through the activation of the TGF-ß/Smad signaling pathway: An in vitro and in vivo study.

The present study aimed to explore the roles of microRNA-21 (miR­21) and the transforming growth factor-ß (TGF-ß)/Smad signaling pathway in the development of peritoneal fibrosis (PF). First, dialysis effluents from 30 patients with PF were collected, and after the establishment of a mouse model of PF, hematoxylin and eosin (H&E) and Masson's staining were used to observe peritoneal tissues, inflammatory cells and blood vessels. High glucose was used to stimulate human peritoneal mesothelial cell lines and these stimulated cells were then transfected with miR­21 inhibitor. Immunofluorescence microscopy was applied for the observation of the transfected cells. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of miR­21, and RT-qPCR and western blot analysis were used to detect the mRNA and protein expression of Zonula occludens-1 (ZO-1), TGF-ß, Smad, vimentin and connective-tissue growth factor (CTGF). The mRNA and protein expression levels TGF-ß, Smad-3, vimentin and CTGF were elevated, while ZO-1 mRNA and protein expression was decreased with the prolonged duration of dialysis treatment in the patients with PF. The experiments using the mouse model of PF revealed that the peritoneal connective tissue was thickened, while the numbers of inflammatory cells and blood vessels were increased. The expression levels of miR­21, and the mRNA and protein expression levels of TGF-ß, Smad-3, vimentin and CTGF were increased over time, whereas the mRNA and protein expression levels ZO-1 constantly decreased in the mice in the experimental group. Moreoever, the expression of miR­21 positively correlated with the expression levels of TGF-ß, Smad-3, vimentin and CTGF, while it negatively correlated with the expression of ZO-1. The results of H&E and Masson's staining revealed that miR­21 expression was associated with the degree of PF. These findings thus indicate that miR­21 promotes the progression of PF through the activation of the TGF-ß/Smad signaling pathway.

Silencing of IRF3 alleviates chronic neuropathic pain following chronic constriction injury.

Interferon regulatory factor 3 (IRF3) is a member of IRF family which plays an important role in neuronal survival and neuroprotection. However, the role of IRF3 in neuropathic pain remains unclear. Thus, in this study, we investigated the effect of IRF3 on neuropathic pain in a rat chronic constriction injury (CCI) model. Our results showed that IRF3 was up-regulated in the dorsal root ganglion (DRG) in CCI rats. Intrathecal short-hairpin RNA (shRNA)-IRF3 attenuated mechanical allodynia and thermal hyperalgesia in CCI rats, as well as inhibited the production of TNF-α and IL-1ß in the DRG of CCI rats. Furthermore, we revealed that sh-IRF3 greatly suppressed the expression of p-NF-κB p65 and IκBα degradation in the DRG of CCI rats. In conclusion, our data suggest that knockdown of IRF3 may alleviate neuropathic pain by inhibiting the activation of NF-κB signaling pathway. Therefore, IRF3 may provide an important target for the treatment of neuropathic pain.

Amino acids-incorporated nanoflowers with an intrinsic peroxidase-like activity.

Functional molecules synthesized by self-assembly between inorganic salts and amino acids have attracted much attention in recent years. A simple method is reported here for fabricating hybrid organic-inorganic nanoflowers using copper (II) ions as the inorganic component and natural amino acids as the organic component. The results indicate that the interactions between amino acid and copper ions cause the growth of the nanoflowers composed by C, N, Cu, P and O elements. The Cu ions and Cu(AA)n complexes containing Cu-O bond are present in the nanoflowers. The nanoflowers have flower-like porous structure dominated by the R groups of amino acids with high surface-to-volume ratios, which is beneficial for exerting its peroxidase-like activity depending on Fenton-like reaction mechanism with ABTS and Rhodamine B as the substrates. It is expected that the nanoflowers hold great promise as enzyme mimics for application in the field of biosensor, bioanalysis and biocatalysis.

[Effects of all-trans retinoic acid on the expression of TGF-beta 1 and COL-I in rat model of peritoneal dialysis].

OBJECTIVE: To investigate the effects of all-trans retinoic acid (ATRA) on the expression of transforming growth factor-beta 1 (TGF-beta 1) and collagen I (COL-I )in rat model of peritoneal dialysis, which may relate to the prevention peritoneal fibrosis. METHODS: Peritoneal dialysis model was established in rats, and then the rats were given ATRA 2 mg/kg (small dose group) or 5 mg/kg (large dose group) by the way of intraperitoneal injection once a day. All the rats were sacrificed on day 28. TGF-beta 1 and COL-I protein expression of peritoneum were measured by immunohistochemistry. TGF-beta 1 mRNA expression were examined with real time polymerase chain reaction (RT-PCR). RESULTS: Masson stain showed that the peritoneum thickness was significantly increased in the rats model, and collagen deposition was evident in the thickened submesothelial compact zone. With the treatment of ATRA, either in small or large dose, pathological changes were significantly lessened. The expression of TGF-beta 1 and COL-I of peritoneum was increased significantly in the rats model, but the levels in the two ATRA treated groups were lower than those of the untreated group. CONCLUSION: ATRA could decrease the experession of TGF-beta 1 and COL-I in peritoneum and delay the progression of peritoneal fibrosis.

[Effect of rosiglitazone on integrin beta1 expression and apoptosis of proximal tubular cell exposed to high glucose].

OBJECTIVE: To evaluate the effect of rosiglitazone (ROS) on integrin beta1 expression and apoptosis of proximal tubular cell exposed to high glucose. METHODS: The proximal tubular cells of rats were cultured in vitro and divided into 6 groups: control group, normal glucose (5 mmol/L) group, high glucose (30 mmol/L) group, only ROS (10 micromol/L) group, glucose (30 mmol/L)+ROS (5 micromol/L) group, and glucose (30 mmol/L) +ROS (10 micromol/L) group. The cells were cultured for 48 hrs and the integrin beta1 expressions were detected by Western blot and RT-PCR; The apoptosis was evaluated by flow cytometry after the cells were cultured for 24, 48, 72 hrs. RESULTS: The expressions of integrin beta1 protein and mRNA of high glucose (30 mmol/L) group were significantly increased as compared with control group and normal glucose (5 mmol/L) group. The integrin beta1 of glucose (30 mmol/L)+ROS (5 or 10 micromol/L) group was decreased as compared with high glucose group (P < 0.05). Flow cytometry demonstrated the decreased apoptosis to be with time-dependence. CONCLUSION: ROS can strikingly inhibit the expression of integrin beta1 in proximal tubular cells exposed to high glucose, with a marked dose-dependent manner. The effect of ROS on cell apoptosis may be relevant to integrin beta1.